SBIR Phase I: Next-Generation Capillary Electrophoresis with Mass Spectrometry for Biopharmaceutical and Biomedical Applications

这项小型企业创新研究（SBIR）项目的更广泛影响/商业潜力将是开发一种强大的毛细管电泳仪器，用于生物医学研究、生物制药开发、临床诊断和系统生物学。毛细管电泳（CE）是一种强大的液体分离技术，在生物分析方面具有显著的潜力。它是完成人类基因组计划的主力工具，在一定程度上导致了DNA测序市场的出现。具有光学和质谱检测的CE系统将使人们能够更深入地了解控制生化过程的分子动力学，如蛋白质表达、解除管制以及与其他生化物种的相互作用，这可能会在药物开发、早期诊断和致命人类疾病（如癌症、糖尿病和神经退行性疾病）的治疗方面取得突破。该提议的技术将通过提高产品表征的效率来提高质量，从而为生物制药药物开发公司带来好处。它也将允许有效的常规蛋白质组学分析和其他生物分析应用。这个SBIR一期项目的智力优势是开发下一代毛细管电泳与电喷雾电离质谱（CE-ESI-MS）用于蛋白质表征。CE-ESI-MS可以高效分离和表征几种生化物质，包括蛋白质。然而，由于缺乏健壮的、易于使用的仪器，利用CE-ESI-MS的内在优势一直很困难，这使得常规应用程序很少采用CE-ESI-MS。本研究将开发和评估一种新型的CE-ESI-MS蛋白质表征仪器。拟议的工具将采用原始创新来克服现有技术的限制并超越其能力。将开发一种新型的流体纳米孔，用于样品和缓冲操作，以允许从1微升或更少的样品体积进行分析。纳米孔将采用全自动设计，减少操作人员的干预。这将是有用的样本有限的应用，如在临床分析。此外，该仪器将利用混合电喷雾电离接口进行同时光学和质谱检测，这是现有技术无法提供的。将CE的超高效率与光学检测和质谱分子鉴定的强大定量特性相结合，将成为生物分析的有力工具。这一组合将代表着对最先进技术的重大进步，并增强了在生物样品中询问蛋白质及其生化功能的手段。提出的创新将显著提高CE-ESI-MS的鲁棒性和能力，用于快速、高效和深入的生物分析。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) project will be the development of a robust capillary electrophoresis instrument with mass spectrometry detection for biomedical research, biopharmaceutical development, clinical diagnostics, and system biology. Capillary electrophoresis (CE) is a powerful liquid separation technology with remarkable potentials for bioanalysis. It was the workhorse tool for the completion of human genome project that, in part, led to the DNA sequencing market. A CE system with both optical and mass spectrometry detection will enable a deeper understanding of molecular dynamics governing biochemical processes, such as protein expression, deregulation, and interaction with other biochemical species, which could lead to breakthroughs in drug development, early diagnosis, and treatments for deadly human diseases such as cancer, diabetics, and neurodegenerative disorders. The proposed technology will offer benefits to biopharmaceutical drug development companies by increasing the efficiency of product characterization for quality improvement. It also will allow efficient routine proteomics analysis and other bioanalytical applications.The intellectual merit of this SBIR Phase I project is to develop a next-generation capillary electrophoresis with electrospray ionization mass spectrometry (CE-ESI-MS) for protein characterization. CE-ESI-MS allows high efficiency separation and characterization of several biochemical species including proteins. However, leveraging the intrinsic advantages of CE-ESI-MS has been difficult due to lack of robust, easy-to-use instrumentation, which has made adoption for routine applications uncommon. This research will develop and evaluate a novel CE-ESI-MS instrument for protein characterization. The proposed instrument will employ original innovations to overcome the limitations of the incumbent technologies and exceed their capabilities. A novel fluidic nanoport for sample and buffer manipulation will be developed to allow analysis from 1 microL or less sample volume. The nanoport will employ a fully automated design with less operator intervention. This will be useful for sample-limited applications such as in clinical analysis. In addition, the instrument will utilize a hybrid electrospray ionization interface for simultaneous optical and MS detection, which no existing technology offers. Combining CE's ultrahigh efficiency with robust quantitative characteristics of optical detection and the molecular identification of mass spectrometry will be a powerful tool for bioanalysis. This combination will represent a significant advancement over the state-of-the-art and enhance the means of interrogating proteins and their biochemical functions in biological samples. The proposed innovations will significantly enhance the robustness and capabilities of CE-ESI-MS for fast, efficient, and deep bioanalysis.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.